Nicotinic acetylcholine receptor α1 promotes calpain-1 activation and macrophage inflammation in hypercholesterolemic nephropathy

Lab Invest. 2011 Jan;91(1):106-23. doi: 10.1038/labinvest.2010.135. Epub 2010 Jul 26.

Abstract

The nicotinic acetylcholine receptor α1 (nAChRα1) was investigated as a potential proinflammatory molecule in the kidney, given a recent report that it is an alternative urokinase plasminogen activator (uPA) receptor, in addition to the classical receptor uPAR. Two animal models and in vitro monocyte studies were involved: (1) In an ApoE(-/-) mouse model of chronic kidney disease, glomerular-resident cells and monocytes/macrophages were identified as the primary cell types that express nAChRα1 during hypercholesterolemia/uninephrectomy-induced nephropathy. Silencing of the nAChRα1 gene for 4 months (6 months on Western diet) prevented the increases in renal monocyte chemoattractant protein-1 and osteopontin expression levels and F4/80+ macrophage infiltration compared with the nonsilenced mice. These changes were associated with significantly reduced transforming growth factor-β1 mRNA (50% decrease) and α smooth muscle actin-positive (αSMA+) myofibroblasts (90% decrease), better glomerular and tubular basement membranes (GBM/TBM) preservation (threefold less disintegration), and better renal function preservation (serum creatinine 40% lower) in the nAChRα1-silenced mice. The nAChRα1 silencing was also associated with significantly reduced renal tissue calcium deposition (78% decrease) and calpain-1 (but not calpain-2) activation (70% decrease). (2) The nAChRα1 was expressed in vitro by mouse monocyte cell line WEHI-274.1. The silencing of nAChRα1 significantly reduced both calpain-1 and -2 activities, and reduced the degradation of the calpain substrate talin. (3) To further explore the role of calpain-1 activity in hypercholesterolemic nephropathy, disease severities were compared in CAST(-/-)ApoE(-/-) (calpain overactive) mice and ApoE(-/-) mice fed with Western diet for 10 months (n=12). Macrophages were the main cell type of renal calpain-1 production in the model. The number of renal F4/80+ macrophages was 10-fold higher in the CAST(-/-)ApoE(-/-) mice (P<0.05), and was associated with a significantly higher level of αSMA+ cells, increased GBM/TBM destruction, and higher serum creatinine levels. Our studies suggest that the receptor nAChRα1 is an important regulator of calpain-1 activation and inflammation in the chronic hypercholesterolemic nephropathy. This new proinflammatory pathway may also be relevant to other disorders beyond hyperlipidemic nephropathy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics
  • Animals
  • Antigens, Differentiation / metabolism
  • Apolipoproteins E / genetics
  • Apolipoproteins E / metabolism
  • Blotting, Northern
  • Blotting, Western
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / metabolism
  • Calpain / metabolism*
  • Cell Line
  • Female
  • Hypercholesterolemia / complications
  • Inflammation / genetics
  • Inflammation / metabolism*
  • Inflammation / pathology
  • Kidney / metabolism
  • Kidney / pathology
  • Kidney Diseases / etiology
  • Kidney Diseases / genetics
  • Kidney Diseases / metabolism*
  • Macrophages / metabolism*
  • Macrophages / pathology
  • Male
  • Mice
  • Mice, 129 Strain
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Monocytes / metabolism
  • Nephrectomy / adverse effects
  • RNA Interference
  • Receptors, Nicotinic / genetics
  • Receptors, Nicotinic / metabolism*
  • Transforming Growth Factor beta1 / genetics

Substances

  • Actins
  • Antigens, Differentiation
  • Apolipoproteins E
  • CHRNA1 protein, mouse
  • Calcium-Binding Proteins
  • Receptors, Nicotinic
  • Transforming Growth Factor beta1
  • monocyte-macrophage differentiation antigen
  • calpastatin
  • Calpain
  • Capn1 protein, mouse